A study on screening and antitumor effect of CD55-specific ligand peptide in cervical cancer cells

Abstract

BackgroundTo improve the targeting ability of antitumor drugs, we identified the antigens with high expression on the surface of tumor cells associated with tumor escape, such as the complement regulatory protein CD55 molecule, which is also known as the decay accelerating factor. In this study, phage display technology was used to screen and identify CD55-specific ligand peptide (CD55sp) bound to CD55 molecule on the surface of cervical cancer HeLa cells. We then explored the role of this peptide in inhibiting the growth of cervical cancer cells in vitro. Our characterization of CD55sp will provide implication for tumor target therapy.MethodsThe phage bound to the surface of HeLa cells were isolated by phage display technology. Positive phage clones were identified by ELISA. Phage was then amplified and determined by agarose gel electrophoresis after monoclonal DNA extraction. DNA sequencing and bioinformatical analysis were conducted to obtain specific ligand peptides. Flow cytometry and immunofluorescence were used to measure the expression of CD55 molecule on the surface of tumor and normal cells. Subsequently, the effects of CD55sp on the proliferation and apoptosis of HeLa and SiHa cells were determined by Cell Counting Kit-8 (CCK-8), flow cytometry, and TUNEL assay, respectively. The morphology of apoptotic cells was examined by electron microscope. The distribution of Cleaved caspase-3 was detected by immunofluorescence. The expression of bcl-2 and Cleaved caspase-3 were determined by Western blot.ResultsThe results showed that the peptide (QVNGLGERSQQM) can bind to the CD55 molecule on the surface of cervical cancer HeLa and SiHa cells as a ligand peptide. It can also effectively inhibit the proliferation of cervical cancer cells and induce cell apoptosis.ConclusionThis study demonstrates that CD55sp screened by phage display technology plays a strong antitumor role.

Clinical value of CD97 and CD55 levels in the differential diagnosis of tuberculous and malignant pleural effusions

“…Our findings show that median CD55 levels in the MPE group were significantly higher than the TPE group, and levels were also inconsistent with other inflammatory factors. These data agree with other studies showing that CD55 contributes to the progression of various cancers [17,32–35] . CD55 may also affect the progress of malignant pleural effusions through complement regulation, however, this conclusion must be confirmed in larger, sample size studies.…”

“…Physiologically, CD55 expression protects host cells from pathogenic microorganisms [31] . As one of the key membrane bound complement regulatory proteins, CD55 is crucial for cancer progression, including lung cancer [17,32–35] . Our findings show that median CD55 levels in the MPE group were significantly higher than the TPE group, and levels were also inconsistent with other inflammatory factors.…”

“…These data agree with other studies showing that CD55 contributes to the progression of various cancers. [17,[32][33][34][35] CD55 may also affect the progress of malignant pleural effusions through complement regulation, however, this conclusion must be confirmed in larger, sample size studies. Pearson correlation analyses showed that CD97 was negatively correlated with CD55 in the MPE group, which does not agree with other studies, for example, Wu et al, [36] showed that CD97 expression was associated with its ligand CD55 in pancreatic cancer, and He et al, [37] also concluded the same for pancreatic cancer.…”

Identification of biomarkers in macrophages of atherosclerosis by microarray analysis

“…CD55 encodes glycoproteins involved in the regulation of complement cascades, which bind to complement proteins and accelerate their own decay, a process that disrupts complement cascades and avoids killing host cells [25]. According to our results, CD55 expression in AS-macrophage samples was significantly lower than that in non-AS tissues.…”

Generation of Single‐Chain Variable Fragment (scFv) Libraries for Use in Phage Display

“…The resulting molecule retains antigen recognition capability and can circumvent the need for hybridomas and immunization (Ahmad et al., 2012). These antibody fragments have substantial clinical potential as delivery vehicles for therapeutic agents targeting tumor‐specific antigens (Liu et al., 2018; Sujjitjoon et al., 2020), as warheads on CAR‐T cells (Sujjitjoon et al., 2020; Zi, Zhao, Wang, Ma, & Wei, 2020), as therapeutic gene delivery agents in cancer (Ahmad et al., 2012; Bakhshinejad, Karimi, & Sadeghizadeh, 2014), and as diagnostic tools and therapeutic agents for autoimmune diseases and neurological disorders (Bazan, Całkosiński, & Gamian, 2012). Their advantages, particularly in cancer treatment, include reduced immunogenicity, better tumor penetration, lower retention times in non‐target tissues (Ahmad et al., 2012), and overall improved therapeutic efficacy of antitumor drugs (Liu et al., 2018).…”